Abstract
By combining low-pressure size-exclusion chromatography with analytical polyacrylamide gel electrophoresis, three stable electrophoretic fractions A, B, and C + D were isolated from soil humic acids (HAs) of chernozem and IHSS standard HA 1S102H. The fractions differ in electrophoretic mobility and molecular size (MS); MSA > MSB > MSC + D. The initial HAs and their fractions are assayed for their biological activity (the ability to stimulate or inhibit the growth of primary roots of 3-day-old radish seedlings) at concentrations of 1, 10–3, and 10–4 mg/L. A statistically significant stimulatory effect (p < 0.05) is observed for both HAs and their fractions A and B (with largest MSs) at an ultralow concentration of 10–3 mg/L. At this concentration, the fraction C + D (with smallest MS) from both HA preparations inhibits the germination of radish seeds. Analysis of the structural characteristics suggests that the main factor determining the growth stimulation is the presence of aliphatic components in HAs and their fractions rather than their MS, whereas their aromatic components inhibit the plant growth.
Similar content being viewed by others
REFERENCES
D. P. Abros’kin, M. Fuentes, J. M. Garcia-Mina, O. I. Klyain, S. V. Senik, D. S. Volkov, I. V. Perminova, and N. A. Kulikova, “The effect of humic acids and their complexes with iron on the functional status of plants grown under iron deficiency,” Eurasian Soil Sci. 49, 1099–1108 (2016). https://doi.org/10.1134/S1064229316100021
S. Guminskii, “The mechanism and conditions of the physiological action of humic substances on the plants,” Pochvovedenie, No. 12, 72–78 (1957).
K. S. Ibragimov and A. D. Fokin, “Absorption of the water-soluble fraction of humic substances by plants from podzolic soil,” Izv. Timiryazevsk. S-kh. Akad., No. 5, 74–80 (1985).
M. M. Kononova, Soil Organic Matter (Academy of Sciences of the Soviet Union, Moscow, 1963) [in Russian].
M. M. Kononova and N. A. Pankova, “The impact of humic substances on the growth and development of plants,” Dokl. Akad. Nauk SSSR 23, 1069–1071 (1950).
O. A. Trubetskoi, S. V. Patsaeva, and O. E. Trubetskaya, “Photochemical degradation of organic pollutants in solutions of soil humic acids,” Eurasian Soil Sci. 52, 1075–1080 (2019). https://doi.org/10.1134/S1064229319090102
O. A. Trubetskoi and O. E. Trubetskaya, “Reversed-phase high-performance liquid chromatography of the stable electrophoretic fractions of soil humic acids,” Eurasian Soil Sci. 48, 148–156 (2015).
O. A. Trubetskoi and O. E. Trubetskaya, “Three-dimensional fluorescence analysis of chernozem humic acids and their electrophoretic fractions,” Eurasian Soil Sci. 50, 1018–1024 (2017).
O. A. Trubetskoi, O. E. Trubetskaya, and C. Saiz-Jimenez, “The study of stable electrophoretic fractions of humic acids by pyrolytic gas chromatography-mass spectrometry,” Eurasian Soil Sci. 38, 1183–1189 (2005).
L. A. Khristeva, “The role of humic acids and other organic substances in the nutrition of higher plants, Pochvovedenie, No. 10, 46–59 (1953).
L. A. Khristeva, “The stimulating effect of humic acid on the growth of higher plants: the nature of this phenomenon,” in Humic Fertilizers: Theory and Practical Use (Kharkov State Univ., Kharkov, 1957), pp. 75–93.
L. A. Khristeva, “Physiological function of humic acid in the metabolism of higher plants,” in Humic Fertilizers: Theory and Practical Use (Kharkov State Univ., Kharkov, 1957), pp. 95–108.
L. A. Khristeva, A. E. Pshenichnyi, and L. R. Pivovarov, “Influence of humic acid on the viability of higher plants in different conditions of mineral nutrition,” in Humic Fertilizers: Theory and Practical Use (Kharkov State Univ., Kharkov, 1957), pp. 109–126.
I. I. Yarchuk, A. E. Pshenichnyi, P. D. Globin, G. V. Kukharevskii, F. S. Kuz’ko, et al., “The efficiency of humic fertilizers,” in Humic Fertilizers: Theory and Practical Use (Kharkov State Univ., Kharkov, 1957), pp. 237–374.
A. Albuzio, S. Nardi, and G. Dell’Anola, “Effect of humic fractions of different molecular size on the development of oat seedlings grown in varied nutritional conditions,” in Humic Substances in the Global Environment and Implication on Human Health, Ed. by N. Senesi and T. M. Miano (Elsevier, Amsterdam, 1994), pp. 199–204.
A. Albusio, S. Nardi, and A. Gulli, “Plant growth regulation activity of small molecular size humic fractions,” Sci. Total Environ. 81–82, 671–674 (1989).
N. Q. Aranson, C. A. Edvards, S. Lee, and R. Byrne, “Effects of humic acids from vermicomposts on plant growth,” Eur. J. Soil Biol. 42, S65–S69 (2006).
J. H. A. Butler and J. N. Ladd, “Importance of the molecular weight of humic and fulvic acids in determining their effects on protease activity,” Soil Biol. Biochem. 3, 249–257 (1971).
W. B. Bottomley, “Some accessory factors in plant growth and nutrition,” Proc. R. Soc. Lond. 88, 237–247 (1914).
L. P. Canellas, D. B. Zandonadi, J. G. Busato, M. A. Baldotto, M. L. Simões, L. Martin-Neto, A. Façanha, R. Spaccini, and A. Piccolo, “Bioactivity and chemical characteristics of humic acids from tropical soils sequence,” Soil Sci. 173, 624–637 (2008).
L. P. Canellas, L. B. Dobbss, A. L. Oliveira, J. G. Chagas, N. O. Aguiar, V. M. Rumjanek, E. H. Novotny, F. L. Olivares, R. Spaccini, and A. Piccolo, “Chemical properties of humic matter as related to induction of plant lateral roots,” Eur. J. Soil Sci. 63, 315–324 (2012).
L. P. Canellas, A. Piccolo, L. B. Dobbss, R. Spaccini, F. L. Olivares, D. B. Zandonadi, and A. R. Façanha, “Chemical composition and bioactivity properties of size-fractions separated from a vermicompost humic acid,” Chemosphere 78, 457–466 (2010).
L. Cavani, C. Ciavatta, O. E. Trubetskaya, O. I. Reznikova, G. V. Afanas’eva, and O. A. Trubetskoj, “Capillary zone electrophoresis of soil humic acid fractions obtained by coupling size-exclusion chromatography and polyacrylamide gel electrophoresis,” J. Chromatogr. A 983, 263–270 (2003).
Y. Chen and T. Aviad, “Effect of humic substances on plant growth,” in Humic Substances in Soil and Crop Science: Selected Readings, Ed. by P. MacCarthy, (Wiley, Chichester, 1990), pp. 161–186.
X. Chen, M. Kou, Z. Tang, A. Zhang, H. Li, and M. Wei, “Responses of root physiological characteristics and yield of sweet potato to humic acid urea fertilizer,” PLoS One 12 (12), e0189715 (2017). https://doi.org/10.1371/journal.pone.0189715
C. Dell’Amico, G. Masciandaro, A. Ganni, B. Ceccanti, C. Garcia, T. Hernandez, and F. Costa, “Effects of specific humic fractions on plant growth,” in Humic Substances in the Global Environment and Implication on Human Health, Ed. by N. Senesi and T.M. Miano (Elsevier, Amsterdam, 1994), pp. 563–566.
E. Fedoseeva, A. Stepanov, O. Yakimenko, S. Patsaeva, M. Freidkin, D. Khundzhua, and V. Terekhova, “Biodegradation of humic substances by microscopic filamentous fungi: chromatographic and spectroscopic proxies,” J. Soils Sediments 19, 2676–2687 (2019).
W. Flaig, “Zur chemie der Huminsäuren und über die physiologische wirkung von modellsubastanzen von Huminsäuren,” Arzneim. Forsch. 4, 402 (1954).
A. C. García, L. G. Ambrosio de Souza, M. G. Pereira, R. N. Castro, J. M. García-Mina, E. Zonta, F. J. G. Lisboa, and R. L. L. Berbara, “Structure-property-function relationship in humic substances to explain the biological activity in plants,” Sci. Rep. 6, 20798 (2016). https://doi.org/10.1038/srep20798
R. D. Hartly and D. S. Whitehead, “Phenolic acids in soils and their influence on plant growth and soil microbial processes,” in Soil Organic Matter and Biological Activity, Ed. by D. Vaughan, (Springer-Verlag, Dordrecht, 1985), Ch. 3, pp. 109–149.
O. I. Klein, E. P. Isakova, Yu. I. Deryabina, N. A. Kulikova, G. A. Badun, M. G. Chernysheva, E. V. Stepanova, and O. V. Koroleva, “Humic substances enhance growth and respiration in the basidiomycetes Trametes maxima under carbon limited conditions,” J. Chem. Ecol. 40, 643–652 (2014).
D. Martinez-Balmori, R. Spaccini, N. Oliveira Aguiar, E. H. Novotny, F. Lopes Olivares, and L. P. Canellas, “Molecular characteristics of humic acids isolated from vermicomposts and their relationship to bioactivity,” J. Agric. Food Chem. 62 (47), 11412–11419 (2014). https://doi.org/10.1021/jf504629c
A. Muscolo, M. Sidari, E. Attinà, O. Francioso, V. Tugnoli, and S. Nardi, “Biological activity of humic substances is related to the chemical structure,” Soil Sci. Soc. Am. J. 71, 75–85 (2007).
A. Muscolo, M. Sidari, and J. A. T. da Silva, “Biological effects of water-soluble soil phenol and humic extract on plant system,” Acta Physiol. Plant. 35 (2), 309–320 (2013). https://doi.org/10.1007/s11738-012-1065-0
S. Nardi, G. Concheri, and G. Dell’Agnola, “Biological activity of humus,” in Humic Substances in Terrestrial Ecosystems, Ed. by A. Piccolo (Elsevier, Amsterdam, 1996), Ch. 9, pp. 361–406.
S. Nardi, D. Pizzeghello, M. Schiavon, and A. Ertani, “Plant biostimulants: physiological responses induced by protein hydrolyzed-bazed products and humic substances in plant metabolism,” Sci. Agric. 73 (1), 18–23 (2016). https://doi.org/10.1590/0103-9016-2015-0006
S. Nardi, D. Pizzeghello, A. Muscolo, and A. Vianello, “Physiological effects of humic substances on higher plants,” Soil Biol. Biochem. 34, 1527–1536 (2002).
S. Nardi, M. R. Panuccino, M. R. Abenavoli, and A. Muscolo, “Auxin-like effect of humic substances extracted from faeces of Allolobophora caliginosa and A. rosea,” Soil Biol. Biochem. 26 (10), 1341–1346 (1994).
A. Piccolo, S. Nardi, and G. Concher, “Structural characteristics of humic substances as related to nitrate uptake and growth regulation in plant systems,” Soil Biol. Biochem. 24 (4), 373–380 (1992.
S. Prat and F. Pospišil, “Humic acids with C14,” Biol. Plant. 1, 71–81 (1959).
C. Richard, O. Trubetskaya, O. Trubetskoj, O. Reznikova, G. Afanas’eva, J. P. Aguer, and G. Guyot, “Key role of the low molecular size fraction of soil humic acids for fluorescence and photoinductive activity,” Environ. Sci. Technol. 38, 2052–2057 (2004).
D. Savy, L. Canellas, G. Vinci, and V. Cozzolino, “Humic-like water-soluble lignins from giant reed (Arundo donax L.) display hormone-like activity on plant growth,” J. Plant Growth Regul. 36 (4), 995–1001 (2017).
Z. Sladky, “The application of extracted humus substances to overground parts of plants,” Biol. Plant. 1, 199–204 (1959).
S. M. Thomas, G. N. Thorne, and I. Pearman, “Effect of nitrogen on growth, yield and photorespiratory activity in spring wheat,” Ann. Bot. 42, 827–837 (1978).
S. Trevisan, O. Francioso, S. Quaggiotti, and S. Nardi, “Humic substances biological activity at the plant-soil interface,” Plant Signaling Behav. 5–6, 635–643 (2010).
O. E. Trubetskaya, O. I. Reznikova, G. V. Afanas’eva, L. F. Markova, T. A. Muranova, and O. A. Trubetskoj, “Amino acids distribution in soil humic acids fractionated by tandem size exclusion chromatography-polyacrylamide gel electrophoresis,” Environ. Int. 24, 573–581 (1998).
O. A. Trubetskoj, P. G. Hatcher, and O. E. Trubetskaya, “1H-NMR and 13C-NMR spectroscopy of chernozem soil humic acid fractionated by combined size-exclusion chromatography and electrophoresis,” Chem. Ecol. 26, 315–325 (2010).
O. A. Trubetskoj, O. E. Trubetskaya, G. V. Afanas’eva, O. I. Reznikova, and C. Sáiz-Jiménez, “Polyacrylamide gel electrophoresis of soil humic acid fractionated by size-exclusion chromatography and ultrafiltration,” J. Chromatogr. A 767, 285–292 (1997).
Z. Varanini, R. Pinton, M. G. De Biasi, S. Astolfi, and A. Maggioni, “Low molecular weight humic substances stimulate H+-ATPase activity of plasma membrane vesicles isolated from oat (Avena sativa L.) roots,” Plant Soil 153, 61–69 (1993).
D. Vaughan, “The stimulation of invertase development in aseptic storage tissue slices by humic acid,” Soil Biol. Biochem. 1, 15–28 (1967).
D. Vaughan and R. E. Malcolm, “Influence of humic substances on growth and physiological processes,” in Soil Organic Matter and Biological Activity, Ed. by D. Vaughan, (Springer-Verlag, Dordrecht, 1985), Ch. 1, pp. 37–75.
D. Vaughan, R. E. Malcolm, and B. G. Ord, “Influence of humic substances on biochemical processes in plants,” in Soil Organic Matter and Biological Activity, Ed. by D. Vaughan, (Springer-Verlag, Dordrecht, 1985), Ch. 2, pp. 77–108.
A. Wallqvist and D. G. Covell, “Hydrophobic interactions in aqueous urea solutions with implications for the mechanism of protein denaturation,” J. Am. Chem. Soc. 120, 427–428 (1998).
Funding
The work was supported by the Russian Foundation for Basic Research (projects nos. 18-016-00078a and 19-05-00056a) and budget funding.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
The authors declare that they have no conflict of interest.
Additional information
Translated by G. Chirikova
Rights and permissions
About this article
Cite this article
Trubetskaya, O.E., Trubetskoi, O.A. The Effect of Low Concentrations of Stable Electrophoretic Fractions of Soil Humic Acids on Stimulation/Inhibition of the Root Length of Radish Seedlings. Eurasian Soil Sc. 54, 1089–1096 (2021). https://doi.org/10.1134/S1064229321060156
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1134/S1064229321060156